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Incomplete luteal regression after treatment with a single dose of prostaglandin F2α during an Ovsynch protocol decreases fertility to timed artificial insemination (TAI). To increase the proportion of cows with complete luteal regression and subsequently pregnancy per AI (P/AI), an additional treatment with prostaglandin F2α (PGF) 24 h after the first has been recommended. This is associated with additional costs due to additional labor required to administer the second PGF treatment and hormones. The objective of this study was to develop a stochastic partial budget model to estimate the economic impact of an additional PGF treatment on d 8 during an Ovsynch protocol. A systematic review of the literature and a meta-analytical assessment was performed to evaluate the effects of adding a second PGF treatment during the Ovsynch protocol on P/AI in lactating dairy cows. Eleven randomized controlled experiments from 10 published manuscripts including 9,432 cows were used. Adding a second PGF treatment during the Ovsynch protocol increased the relative risk for pregnancy (RR = 1.17; 95% CI = 1.11 to 1.23) in lactating dairy cows 32 d after TAI using the fixed effects model. An additional treatment with PGF yielded a 5.73 risk difference in P/ AI (95% CI = 3.79 to 7.68) using a fixed effects model. Revenue was based on the associated improvement in reproductive performance due to an increase in P/AI multiplied by the value of a pregnancy (PGVAL). Median PGVAL was €252 (5% percentile €42 and 95% percentile €623) with a minimum of −€138 and a maximum of €1,198. The median PGVAL varied among the 10 farms ranging from €192 to €462. Using a stochastic simulation model with 10,000 iterations adding a second PGF dose on d 8 during an Ovsynch protocol was more profitable (€7.98/ cow; 5% percentile €0.25 and 95% percentile €30.31) compared with a single PGF administration on d 7. The median return on investment was €2.17. In conclusion, there was a clear benefit of an additional PGF treatment during the Ovsynch protocol on P/AI (+5.7 percentage units). Despite additional costs for hormones and labor an additional treatment with prostaglandin F2α on d 8 was more profitable in 95% of all scenarios due to the associated increase in fertility.